The invention provides novel methods for isolating microvesicles from a biological sample and for extracting nucleic acids from the microvesicles.

The invention provides novel methods for isolating microvesicles from a biological sample and for extracting nucleic acids from the microvesicles.

A membrane, method and system are disclosed for rapid, sensitive and precise detection of an agent suspected of being present in a sample. The agent may be a cell or microorganism, e.g., a single pathogenic bacteria, and the sample may be small, e.g., milliliters of unprocessed environmental water. The sample is processed by filtering it through an asymmetric membrane having multiple layers. One layer has microchannels for capturing the agent and another layer has nanochannels for passing particles smaller than the agent. Amplification reagents, such as loop-mediated isothermal amplification (LAMP) reagents, are load onto membrane so that the microchannels act as nanoreactors, creating quantifiable amplicons within the pores on the exposed surface of the membrane in response to captured agent. The amplicons may be imaged and counted using a fluorescent camera. The membrane is capable of agent capture, concentration, purification, partition, lysis and digital LAMP without off-membrane sample treatments.

A membrane, method and system are disclosed for rapid, sensitive and precise detection of an agent suspected of being present in a sample. The agent may be a cell or microorganism, e.g., a single pathogenic bacteria, and the sample may be small, e.g., milliliters of unprocessed environmental water. The sample is processed by filtering it through an asymmetric membrane having multiple layers. One layer has microchannels for capturing the agent and another layer has nanochannels for passing particles smaller than the agent. Amplification reagents, such as loop-mediated isothermal amplification (LAMP) reagents, are load onto membrane so that the microchannels act as nanoreactors, creating quantifiable amplicons within the pores on the exposed surface of the membrane in response to captured agent. The amplicons may be imaged and counted using a fluorescent camera. The membrane is capable of agent capture, concentration, purification, partition, lysis and digital LAMP without off-membrane sample treatments.

The present invention relates, in part, to methods for large-scale purification of mRNA. The method includes, at least, steps of forming an mRNA slurry, stirring the slurry, and vacuum or pressure filtering the slurry.

Multivalent binding compositions including a particle-nucleotide conjugate having a plurality of copies of a nucleotide attached to the particle are described. The multivalent binding compositions allow one to localize detectable signals to active regions of biochemical interaction, e.g., sites of protein-protein interaction, protein-nucleic acid interaction, nucleic acid hybridization, or enzymatic reaction, and can be used to identify sites of base incorporation in elongating nucleic acid chains during polymerase reactions and to provide improved base discrimination for sequencing and array based applications.

Multivalent binding compositions including a particle-nucleotide conjugate having a plurality of copies of a nucleotide attached to the particle are described. The multivalent binding compositions allow one to localize detectable signals to active regions of biochemical interaction, e.g., sites of protein-protein interaction, protein-nucleic acid interaction, nucleic acid hybridization, or enzymatic reaction, and can be used to identify sites of base incorporation in elongating nucleic acid chains during polymerase reactions and to provide improved base discrimination for sequencing and array based applications.

Multivalent binding compositions including a particle-nucleotide conjugate having a plurality of copies of a nucleotide attached to the particle are described. The multivalent binding compositions allow one to localize detectable signals to active regions of biochemical interaction, e.g., sites of protein-protein interaction, protein-nucleic acid interaction, nucleic acid hybridization, or enzymatic reaction, and can be used to identify sites of base incorporation in elongating nucleic acid chains during polymerase reactions and to provide improved base discrimination for sequencing and array based applications.

Multivalent binding compositions including a particle-nucleotide conjugate having a plurality of copies of a nucleotide attached to the particle are described. The multivalent binding compositions allow one to localize detectable signals to active regions of biochemical interaction, e.g., sites of protein-protein interaction, protein-nucleic acid interaction, nucleic acid hybridization, or enzymatic reaction, and can be used to identify sites of base incorporation in elongating nucleic acid chains during polymerase reactions and to provide improved base discrimination for sequencing and array based applications.

Methods and materials are disclosed for use in recovering a biopolymer from a solution. In particular, the invention provides methods for extraction and isolation of nucleic acids from biological materials. The nucleic acids can be separated by forming a stable complex with soluble polysaccharide polymers and magnetic particles, in the presence of detergents and solvent. When the particles are magnetically separated out of the solution, the nucleic acids are separated with them. The nucleic acids can subsequently be released and separated from the particles. The nucleic acid preparation is useful for achieving efficient and accurate results in downstream molecular techniques such as quantification, identification of the source of the nucleic acids, and genotyping.